Anti-Folate Binding Protein/FBP antibody [EPR20277] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] - BSA and Azide free (AB235140)

This data was developed using ab221543 the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of formalin fixed paraffin embedded human cervical carcinoma labelling Folate Binding Protein with ab221543 at a concentration of 0.1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32 mins. ab221543 Anti-Folate Binding Protein/FBP antibody [EPR20277] was incubated for 16 mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] - BSA and Azide free (AB235140)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Folate Binding Protein/FBP with ab221543 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051, ready to use). Cytoplasmic staining on human kidney (PMID : 22204844). Counter stained with Hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051, ready to use).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab221543).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] - BSA and Azide free (AB235140)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Folate Binding Protein/FBP with ab221543 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use). Positive staining on rat kidney (PMID : 22204844). Counter stained with Hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab221543).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] - BSA and Azide free (AB235140)

Negative control :

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Folate Binding Protein/FBP with ab221543 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use). No staining on human tonsil (PMID : 22547449). Counter stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab221543).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] - BSA and Azide free (AB235140)

Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling Folate Binding Protein/ FBP with ab221543 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use). Cytoplasmic staining on tumor cells of human lung cancer (PMID : 22204844). Counter stained with Hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab221543).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] - BSA and Azide free (AB235140)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Folate Binding Protein/FBP with ab221543 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use). Positive staining on mouse kidney (PMID : 22204844). Counter stained with Hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab221543).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Folate Binding Protein/FBP antibody [EPR20277] - BSA and Azide free (AB235140)

Immunohistochemical analysis of paraffin-embedded human ovarian cancer tissue labeling Folate Binding Protein / FBP with ab221543 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use). Mainly membranous staining on tumor cells of human ovarian cancer (PMID : 22204844). Counter stained with Hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97501, ready to use).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab221543).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-Folate Binding Protein/FBP antibody [EPR20277] - BSA and Azide free (AB235140)

This data was developed using the same antibody clone in a different buffer formulation (ab221543).

Lanes 1-4 : Merged signal (red and green). Green - ab221543 observed at 36, 38 kDa. Red - loading control, ab7291 observed at 52 kDa.

ab221543 Anti-Folate Binding Protein/FBP antibody [EPR20277] was shown to specifically react with Folate Binding Protein/FBP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264921 (knockout cell lysate ab257270) was used. Wild-type and Folate Binding Protein/FBP knockout samples were subjected to SDS-PAGE. ab221543 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

The molecular weight (36,38kDa) observed were knockout validated and consistent with what has been described in the literature (PMID : 23144806).

All lanes:

Western blot - Anti-Folate Binding Protein/FBP antibody [EPR20277] (<a href='/en-us/products/primary-antibodies/folate-binding-protein-fbp-antibody-epr20277-ab221543'>ab221543</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human FOLR1 (Folate Binding Protein/FBP) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-folr1-folate-binding-protein-fbp-knockout-hela-cell-line-ab264921'>ab264921</a>)

Lane 2:

FOLR1 knockout HeLa cell lysate at 20 µg

Lane 3:

JAR cell lysate at 20 µg

Lane 4:

MDA-MB-231 cell lysate at 20 µg

Predicted band size: 30 kDa

Observed band size: 38 kDa,36 kDa

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